The Subsets Of Immune Cells And Plasma Cytokines In SIVmac239 Infected Rhesus Monkeys By HAART Treatment

CD4+T cell decrease caused by HIV invasion is the root cause of AIDS.In recent years,with the extention of HAART and therapeutic schedule improvemented,the plasma viral load of most HIV infected patients has been well controlled,and the peripheral blood CD4+ T cell count can also be maintained at a normal level.However,with the increase of patients’ age and the prolongation of HAART treatment time,the risk of HIV-infected people suffering from non AIDS related diseases is escalates,and has become one of the important factors affecting the quality of life of HIV infected people.There is more and more evidence that the risk of Non-AIDS-defining diseases is closely related to the level of immune activation and inflammation in infected people.In this regard,we have carried out the following parts of the study.Part Ⅰ:screening and bioinformatics analysis of differential genes between infected and uninfected patients treated with HAART.After HIV infection,long-term incubation and medication lead to complex immune activation and inflammation in the body,which makes it difficult to obtain comprehensive immune activation and inflammation results through limited in vivo and in vitro experiments.Bioinformatics technology can collect and sort out gene detection data of AIDS patients in multiple experiments through database.Regrouping according to the infection strain,infection time,drug administration,gender,age,ethnic background and tissue samples can accurately predict the key direction of experimental verification and the sample size used before the laboratory experiment.In this study,we screened the differential genes of CD4+T lymphocytes,CD8+T lymphocytes,monocytes and PBMC between HIV infected and untreated elite infected with HAART,and studied the main genes involved in lymphocyte immune activation of HIV infected patients treated with HAART by GO,KEGG and PPI network analysis system and the function of these genes.In addition,we also analyzed the differences between HIV infected people and healthy controls at miRNA level.The results showed that HAART could reduce the expression of most inflammatory genes,but also increase the expression of chemoreceptor genes IL-7R and CX3CR1 in CD4+T,NFKBIA genes in CD8+T cells and CXCL10 in monocytes.After HAART treatment,lymphocyte subsets regulate immune activation and inflammatory state through different ways.CD4+T lymphocytes mainly downregulate the regulatory pathway of leukocyte activation and the expression of negative regulatory genes of leukocyte activation.CD8+T cells down regulate innate immunity and activate αβ CD8+T cells.Monocytes can affect the immune activation process of HIV infected people by up regulating cytokine mediated signal pathway,lymphocyte chemoattractant gene,down regulating adaptive immune response,cell defense response and NK cell-mediated immune genes.In addition,HAART treatment can change the expression of tumor related genes such as SERPINA1、S100A9、IFIT2 and CD247,as well as the effects on insulin resistance,neuron apoptosis and other pathways.Part Ⅱ:The expression of cytokines in the plasma of rhesus monkeys infected with SIVmac 239 and its correlation with disease characteristics after HAART treatment.Through the screening of bioinformatics technology,we can get the data of HAART treatment changing genes expression of infected people.Can we detect the corresponding changes of cytokines level which can reflect the inflammatory state in the established experimental animal model?In this study,we used Luminex liquid chip technology to study the plasma concentrations of these cytokines in monkeys infected with SIVmac239 treated by HAART before infection,during chronic infection,under HAART treatment and drug withdrawal,and analyzed the correlation between the changes of these cytokines and disease characteristics.The results showed that the concentration of cytokines in the plasma of rhesus monkeys injected with SIVmac 239 increased in the chronic stage of infection.During HAART treatment,the concentrations of TGF-β1 and IP-10 were consistent with the clinical trend and the correlation with disease characteristics.The plasma concentrations of IL-1β and IL-6 did not decrease with the decrease of virus level,but gradually decreased after halt treatment and basically maintained at the level before HAART treatment.Part III:The changes of immune cell subsets in rhesus monkeys infected with SIVmac239 after HAART and their correlation with disease characteristics.The change of cytokines concentration in plasma can well understand the dynamic of inflammation,but it can not respond well to HAART in the treatment of SIVmac239 infected rhesus monkeys.We can’t systematically know the relationship between immune activation and inflammation.Therefore,the number and proportion of immune cell subsets in rhesus monkeys infected with SIVmac239 treated by HAART were detected by multi-color flow cytometry,and the correlation between these immune cell subsets and the expression level of cytokines was analyzed.The results showed that the subpopulation of CD4+TEMRA,CD4+TCM and CD4+TEM cells with higher differentiation degree decreased in the course of AIDS,while the proportion of CD4+naive T cells with lower differentiation degree increased in the course of AIDS.HAART treatment did not change the overall trend that the proportion of CD8+TEMRA cell subsets continued to decrease and the proportion of CD8+naive T continued to increase after HIV infection.HAART can not effectively restore the level of CD8+TCM cells.PD-I was highly expressed on the surface of CD4+TEMRA and CD8+TEMRA cells,which were in low level on the surface of corresponding naive T cells.HAART treatment reduced the expression of PD-1 on the surface of CD8+T memory cell subpopulation only in the early stage of treatment.On day 112,the proportion of B1 cells in peripheral blood increased,and the proportion of classical monocytes with phagocytic function and activated CD 14+CD169+monocytes in peripheral blood also increased significantly.CD8+CD38+T as an indicator of CD4+T count recovery in peripheral blood was verified in the rhesus monkey model infected with SIVmac 239 treated by HAART.